Purification notesWhole antiserum is fractionated and further purified by anion-exchange chromatography to provide the IgG fraction of antiserum that is essentially free of other rabbit serum proteins.

Target

FunctionResponsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Histone deacetylases act via the formation of large multiprotein complexes. Deacetylates SP proteins, SP1 and SP3, and regulates their function. Component of the BRG1-RB1-HDAC1 complex, which negatively regulates the CREST-mediated transcription in resting neurons. Upon calcium stimulation, HDAC1 is released from the complex and CREBBP is recruited, which facilitates transcriptional activation. Deacetylates TSHZ3 and regulates its transcriptional repressor activity. Deacetylates 'Lys-310' in RELA and thereby inhibits the transcriptional activity of NF-kappa-B.

Tissue specificityUbiquitous, with higher levels in heart, pancreas and testis, and lower levels in kidney and brain.

Post-translationalmodificationsSumoylated on Lys-444 and Lys-476; which promotes enzymatic activity. Desumoylated by SENP1.Phosphorylation on Ser-421 and Ser-423 promotes enzymatic activity and interactions with NuRD and SIN3 complexes.Ubiquitinated by CHFR, leading to its degradation by the proteasome.

Anti-HDAC1 antibody - ChIP Grade images

Immunofluorescence - Anti-HDAC1 antibody - ChIP Grade (ab7028)

HeLa cells were fixed and permeabilized with cold methanol followed by cold methanol:acetone. Fixed cells were stained with 1:200 ab7028. The antibody was developed using Goat Anti-Rabbit IgG, Cy3 conjugate.

ChIP - Anti-HDAC1 antibody - ChIP Grade (ab7028)Sylvain Daujat

Sonicated Chromatin prepared from untreated (UI) or 17beta-estradiol (E2) treated MCF7 cells was subjected to the ChIP procedure with ab7028 to HDAC1 and the immunoprecipitated chromatin was analysed in the proximal region of the estrogen-responsive pS2 promoter (as shown above) and quantified by real-time PCR (values are nomalized over inputs). The primers are designed to follow the nucleosome E (including the Estrogen Responsive Element ERE). 5 µl of ab7028 and 2x10.

ab7028 staining HDAC1 in human primary cardiac cells by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with permeabilization buffer. The sample was incubated with the undiluted primary antibody for 12 hours at 4°C. An Allophycocyanin-conjugated goat polyclonal anti-rabbit IgG (1/1000) was used as the secondary antibody.Gating Strategy: Isotype negative control (white).

HDAC1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to HDAC1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab7028.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 60ka: HDAC1.

Hagelkruys A et al.A single allele of Hdac2 but not Hdac1 is sufficient for normal mouse brain development in the absence of its paralog.Development141:604-16 (2014).
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Read more (PubMed: 24449838) »

Thank you so much for keeping me updated about the situation, and I am so happy to hear that the replacement from a new lot is working well. Please let me know if there is anything else that you need, and have a great week!

Thanks for your call today and letting us know about the trouble with the new lot of ab7028.
I am sending a free of charge vial from a new lot on the order ***, which should arrive tomorrow.
Please keep me updated with the results using...

Thank you for your patience while I have been in touch with the lab regarding the antigen retrieval method for ab7028.
I have heard from the lab that they used 0.1% pronase for 5 - 10 minutes in order to expose the epitope.
Please let me know if...